JP2010241734A - Skin external preparation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To solve such problem that there is concern for causing problems of odor, stability and the like due to the interactions among amino acids or between one amino acid and other amino acids when 15 amino acids are simultaneously used although it has long been aware that 15 kinds of amino acids are simultaneously incorporated into skin external preparations. <P>SOLUTION: It has been found that the combination of five kinds of alanine, proline, arginine, glutamic acid, and glycine or the salts thereof exhibits a high anti-aging effect, and moreover the combination of these five kinds of amino acids with an extract of Casuarina stricta exhibits a higher anti-aging effect. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

    The present invention relates to a skin external preparation containing five kinds of amino acids (alanine, proline, arginine, glutamic acid, glycine, or a salt thereof), and further to a skin external preparation containing an extract of Casuarina Adans.

    Human skin tissue is composed of cell layers such as fibroblasts and epidermal cells, and a dermal matrix such as collagen and hyaluronic acid that supports the cell layers. Further, collagen, hyaluronic acid and the like, which are constituents of the dermal matrix, decrease with aging, and this is considered to be a cause of skin firmness, tarmi, wrinkles and the like.

    In order to improve skin condition such as skin firmness, collagen is sometimes added to cosmetics as a natural moisturizing agent. Such cosmetics improve the keratinous state of the epidermis, and the dermal matrix It does not improve to the state of. In addition, collagen, which is an animal-derived protein, may exhibit strong antigenicity, and when blended with cosmetics, inflammation may occur due to sensitization depending on the user. For this reason, in order to increase the amount of collagen produced by the user's dermal fibroblasts, attempts have been made to add various substances to cosmetics and the like.

    So far, Patent Document 1 discloses that 15 kinds of amino acids are used in combination in an external preparation for skin and have the effect of increasing the amount of collagen produced by human dermal fibroblasts. However, when 15 kinds of amino acids are used in combination, there is a concern that odor problems and stability problems may occur due to the interaction between amino acids or other components, and various active ingredients are searched for and studied for compounding. I came.

For example, in order to obtain an external preparation for skin having anti-aging and improving effects on skin, there is an essence of Ponkan (see Patent Document 2) as a component having an activity of dermal fibroblast activation or proliferation, and is used in combination with an amino acid. With regard to the active ingredient, an external preparation for skin containing a salt of an alkyl ester of oxidized and reduced glutathione and an amino acid (Patent Document 3), whey, and an external preparation containing amino acid (serine, alanine, trimethylglycine) And the like, and with regard to the extract of humus, a moisturizer, cell activator, dermal fibroblast activator, epidermis cell activator, collagen production promoter, antitumor, Production of type IV collagen containing oxidants, anti-aging agents, whitening agents, melanin production inhibitors (Patent Document 5), and asteraceae plants as active ingredients Susumuzai, and basement membrane reinforcing agent (Patent Document 6) have been disclosed.
JP 2007-051087 A JP 2001-131045 A Japanese Patent No. 4002313 JP 2000-178165 A JP 2007-277100 A JP 2008-24622 A

    Patent Document 1 discloses that 15 types of amino acids are used in combination in an external preparation for skin and have an effect of increasing the amount of collagen produced by human dermal fibroblasts. However, when 15 kinds of amino acids are used in combination, there is a concern that an odor problem or a stability problem may occur due to the interaction between amino acids or other components.

    A combination of five types of alanine, proline, arginine, glutamic acid, glycine, or their salts has been found to exert a high anti-aging effect, and further, the combination of these five types of amino acids and Mokumau extract is high. It has been found that it exhibits an anti-aging effect.

    According to the present invention, it has been found that by combining five kinds of amino acids of alanine, proline, arginine, glutamic acid, glycine, or a salt thereof, a high anti-aging effect is exerted, and further, by combining a mocoum extract. A topical skin preparation that exhibits a high anti-aging effect can be provided.

    The five amino acids used in the present invention are preferably alanine, proline, arginine, glutamic acid, glycine, or a salt thereof.

    As the five amino acids used in the present invention, such as alanine, proline, arginine, glutamic acid, glycine, or a salt thereof, it is preferable to use easily available amino acids.

    The humus used in the present invention is a dicotyledonous plant belonging to the genus Casuarinaceae and genus Moumuma, which is an evergreen tree, with about 60 species distributed mainly in Australia, and a few species in Southeast Asia and Polynesia.

    There are no particular restrictions on the area of use when using the pupa, and each part such as leaves, stems, flowers, roots, and whole plants can be used, but leaves and branches are preferably used.

    At the time of extraction, you can use the cockatoo as raw, but considering the extraction efficiency, it is preferable to perform the extraction after processing such as shredding, drying, and pulverization.

    The extraction can be performed by immersing in an arbitrary extraction solvent for a predetermined time. The extraction solvent may be heated as necessary. In order to increase extraction efficiency, stirring or homogenization in an extraction solvent may be performed. The extraction temperature is suitably about 5 ° C. to the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but is suitably about 1 hour to 14 days.

    As an extraction solvent, in addition to water, lower alcohols such as methanol, ethanol, propanol and isopropanol; polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol and glycerin; ethers such as ethyl ether and propyl ether Solvents such as esters such as butyl acetate and ethyl acetate; ketones such as acetone and ethyl methyl ketone can be used, and water and ethanol are preferable. These may be used alone or in combination of any two or more. Saline, phosphate buffer, phosphate buffered saline, and the like may be used. Furthermore, you may use 1 type, or 2 or more types of supercritical liquids and subcritical liquids, such as water, a carbon dioxide, ethylene, propylene, ethanol, methanol, ammonia.

    The extract of the above-mentioned solvent of ummao used in the present invention can be used as it is, but it may be used by leaving it to stand for a certain period of time and aged, or the concentrated and dried product can be used as water or a polar solvent. It can be dissolved again and used. Alternatively, it may be used after performing purification treatment such as decolorization, deodorization, and desalting, and fractionation treatment by column chromatography or the like within a range not impairing these physiological functions. The said extract of plant and shellfish, its processed material, and the fractionation thing can also be freeze-dried after each processing and fractionation, and can also be melt | dissolved and used for a use solvent. It can also be used by encapsulating in vesicles such as liposomes or microcapsules.

    A combination of alanine, proline, arginine, glutamic acid, glycine, or a salt thereof has been found to exert a high anti-aging effect, and further, as a skin external preparation that exerts a high anti-aging effect by combining with Mokumau extract. Can be used.

    A topical skin preparation comprising a combination of five types of amino acids has an effect of activating human dermal fibroblasts and exhibits an excellent effect in preventing and improving skin aging symptoms.

    An external preparation for skin comprising a combination of five types of amino acids has an action of promoting human dermal fibroblast type I collagen production and exhibits an excellent effect in preventing and improving skin aging symptoms.

    An external preparation for skin, which is a combination of five kinds of amino acids and Mokumau extract, has an action of promoting human dermal fibroblast type I collagen production, and exhibits an excellent effect in preventing and improving skin aging symptoms.

    A topical skin preparation that combines five types of amino acids has an effect of improving skin firmness and exhibits an excellent effect in preventing and improving skin aging symptoms.

    A topical skin preparation comprising a combination of 5 kinds of amino acids and Mokumau extract has an effect of improving skin firmness and exhibits an excellent effect in preventing and improving skin aging symptoms.

    Here, the external preparation for skin means all external compositions for external use on the skin or hair, such as cosmetics, quasi-drugs or external pharmaceuticals.

    The dosage form of the external preparation for skin is arbitrary, and can be provided, for example, as a solubilization system such as lotion, a dispersion system such as calamine lotion, or an emulsification system such as cream or emulsion. Furthermore, it can also be provided in various dosage forms such as aerosol form, ointment or poultice filled with propellant.

    Specifically, various cosmetics such as emulsions, creams, lotions, lotions, packs, cosmetic liquids, cleaning agents or makeup cosmetics; liquids, ointments, powders, granules, aerosols, patches or poultices Various forms of cosmetics, quasi-drugs, or external medicines can be exemplified.

    In addition to the five types of amino acids or mokumao extract, the topical skin preparation is usually blended into pharmaceuticals, quasi drugs, skin cosmetics, hair cosmetics, and cleansing agents as needed. Optional ingredients such as water, oily ingredients, humectants, powders, pigments, emulsifiers, solubilizers, gelling agents, cleaning agents, UV absorbers, anti-inflammatory agents, thickeners, pH adjusters, chelating agents, Drugs (medicinal ingredients), fragrances, resins, fungicides, fungicides, antioxidants, alcohols, and the like can be appropriately blended. Furthermore, in the range which does not impair the effect of the present invention, other anti-aging agents, amino acids other than the five amino acids, or other plant extracts can be used in combination.

    The blending amount of the five types of amino acids into the external preparation for skin can be adjusted depending on the type, purpose, etc. From the viewpoints of effects and stability, each amino acid is preferably converted into solid content with respect to the total amount, preferably It is 0.00001-10.0 mass%, More preferably, it is 0.0001-5.0 mass%, More preferably, it is 0.001-5.0 mass%, More preferably, it is 0.001-1. 0.0% by mass.

    The blending amount of the extract of the umbilical extract into the external preparation for skin can be adjusted depending on the type, purpose, etc., but from the viewpoint of effect and stability, the total amount is preferably 0.00001 to 10 in terms of solid content. 0.0 mass%, more preferably 0.0001-5.0 mass%, still more preferably 0.001-5.0 mass%, and still more preferably 0.01-5.0 mass%. is there.

    In the following, preparation examples of the extract of mokumau, test methods for evaluating anti-aging effects, and formulation examples as external preparations for skin will be described in more detail, but the technical scope of the present invention is not limited by these. Absent.

[Extract 1]
To 100 g of dried pulverized leaves and branches, 2.0 kg of a 50% by mass aqueous ethanol solution was added and extracted at room temperature for 2 hours with stirring. The extract was collected by filtration, concentrated under reduced pressure, and lyophilized to obtain extract 1.

    Each effect was evaluated using the said extract. Note that * and ** described in each evaluation result indicate that the significance probability P value in the t-test is less than 5% (P <0.05), and less than 1% significance (P <0.01). ) Is represented by **.

[Example 1]
<Evaluation of human dermal fibroblast activation effect>
Normal human dermal fibroblasts manufactured by Kurabo Industries Co., Ltd. (Kurashiki Boseki Co., Ltd.) were seeded in a 96-well microplate so as to be 2.0 × 10 ⁴ cells per well. As the seeding medium, Dulbecco's modified Eagle medium (DMEM) to which 1% by mass of fetal bovine serum (FBS) was added was used. After culturing for 24 hours, the medium was replaced with 1% by mass FBS-added DMEM medium to which amino acids were added so as to achieve the respective concentrations shown in Table 1, and further cultured for 48 hours. After removing the supernatant, the medium was replaced with a medium containing 400 μg / mL of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide (MTT) and cultured for about 2 hours. Thereafter, formazan produced by the opening of the tetrazolium ring was extracted with 2-propanol, and the absorbance at 550 nm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values. In the evaluation, in addition to the medium containing the sample, 1% by mass FBS-added DMEM medium was used as a control. The evaluation results are shown in Table 1 as relative values when the cell activation effect in the control is 100.

    Evaluation result of synergistic effect of human dermal fibroblast activation using 5 kinds of amino acids

    As is clear from the results in Table 1, a combination of 5 amino acids showed a significant human dermal fibroblast activation effect as compared with those added with amino acids.

[Example 2]
<Anti-aging effect (Evaluation of human dermal fibroblast type I collagen production promoting effect)>
Normal human dermal fibroblasts were seeded in a 96-well microplate at 2.0 × 10 ⁴ cells per well. Dulbecco's modified Eagle medium (DMEM) supplemented with 5% by weight fetal bovine serum (FBS) was used as the seeding medium. After 24 hours, the medium was replaced with a culture solution to which amino acids and / or extract 1 were added so as to achieve the respective concentrations shown in Table 2 in a DMEM medium supplemented with 0.5% by mass FBS, and further cultured for 24 hours. The ELISA method was used for the quantification of type I collagen secreted into the culture supernatant. Finally, 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt ( ABTS) and hydrogen peroxide were added and reacted, and then the absorbance at 405 nm was measured. The amount of protein in each well was measured with a BCA protein assay kit manufactured by PIERCE, and the amount of type I collagen produced per unit protein amount was determined.
The evaluation results are shown in Table 2 as relative values when the type I collagen production amount per unit protein amount in the control with no sample added is defined as 100.

    Evaluation results of synergistic effect of human dermal fibroblast type I collagen production promoting action using 5 kinds of amino acids and Mokumau extract

    From the results shown in Table 2, the combination of 5 types of amino acids has a human dermal fibroblast type I collagen production promoting action, and the addition of mocoum has significant human dermal fibroblast type I collagen production. A promoting action was observed.

    From the above results, it has been found that the addition of 5 kinds of amino acids and further the extract of Mokumaum exhibits an anti-aging effect.

[Example 3]
1. Measurement of elasticity on human skin A measuring instrument Cutometer SEM575 (Courage-Khazaaka slectronic GmbH, Cologene, Germany) using a suction method was used. This is because a probe having a 2 mm circular opening is lightly adhered to a smooth skin surface, and then the skin is sucked into the probe by making the inside of the probe a constant negative pressure, which is displaced when bulging and deforming. The elasticity of the skin is measured by measuring the amount and displacement until the end of suction.
In the test, the elasticity of the skin before use was measured with a Cutometer SEM575 in the range of 3 × 4 cm ² of the forearm, and then 24 μL of each sample aqueous solution was applied within the range. The tension value (R7) value was measured at 15, 60 and 120 minutes after application.
The evaluation results are shown in Table 3.

    Elasticity measurement results on human skin using 5 types of amino acids

有意確率 p＜0.05：* ｐ＜0.01：** （ｔ検定による）

Significance probability p <0.05: * p <0.01: ** (by t-test)

    From the results of Table 3, it was found that the addition of 5 types of amino acids maintained the elasticity of human skin even after 120 minutes and improved skin firmness.

    Then, the formulation example of the skin external preparation which mix | blended five types of amino acids obtained by each said preparation method and / or Mokumao extract is shown.

[Example 4]
Emulsion (1) Squalane 10.0 (mass%)
(2) Methylphenylpolysiloxane 4.0
(3) Hydrogenated palm kernel oil 0.5
(4) Hydrogenated soybean phospholipid 0.1
(5) Polyoxyethylene monostearate
Sorbitan (20E.O.) 1.3
(6) Sorbitan monostearate 1.0
(7) Glycerin 4.0
(8) Methyl paraoxybenzoate 0.1
(9) Carboxyvinyl polymer 0.15
(10) The balance of 100 purified water (11) Alanine 0.04
(12) Proline 0.04
(13) Arginine 0.04
(14) Glutamic acid 0.04
(15) Glycine 0.04
(16) Extract 1 0.05
Production method: The oil phase components (1) to (6) are heated and dissolved at 80 ° C. On the other hand, the aqueous phase components (7) to (15) are dissolved by heating at 80 ° C. The oil phase component is added to this while stirring and uniformly emulsified with a homogenizer. After cooling, add (16) sequentially at 40 ° C. and mix uniformly.

[Example 5]
Lotion (1) Ethanol 15.0 (mass%)
(2) Polyoxyethylene (40E.O.) hydrogenated castor oil 0.3
(3) Fragrance 0.1
(4) Purified water 100 (5) Citric acid 0.02
(6) Sodium citrate 0.1
(7) Glycerin 1.0
(8) Hydroxyethyl cellulose 0.1
(9) Alanine 0.04
(10) Proline 0.04
(11) Arginine 0.04
(12) Glutamic acid 0.04
(13) Glycine 0.04
Production method: (2) and (3) are dissolved in (1). Further, after sequentially adding (4) to (13), the mixture is sufficiently stirred, and (14) is added and mixed uniformly.

[Example 6]
Cream (1) Squalane 10.0 (mass%)
(2) Stearic acid 2.0
(3) Hydrogenated palm kernel oil 0.5
(4) Hydrogenated soybean phospholipid 0.1
(5) Cetanol 3.6
(6) Lipophilic glyceryl monostearate 2.0
(7) Glycerin 10.0
(8) Methyl paraoxybenzoate 0.1
(9) Purified water 100 (10) Alanine 0.04
(11) Proline 0.04
(12) Arginine 0.04
(13) Glutamic acid 0.04
(14) Glycine 0.04
(15) Carboxyvinyl polymer (1% by weight aqueous solution) 15.0
(16) Extract 1 0.05
Production method: The oil phase components (1) to (6) are heated and dissolved at 80 ° C. On the other hand, the aqueous phase components (7) to (14) are dissolved by heating at 80 ° C. The oil phase component is added to this while stirring and uniformly emulsified with a homogenizer. Add (15), stir, cool and add (16) at 40 ° C. and mix uniformly.

[Example 7]
Essence (1) Purified water 100 balance (mass%)
(2) Glycerin 10.0
(3) Sucrose fatty acid ester 1.3
(4) Carboxyvinyl polymer (1% by weight aqueous solution) 17.5
(5) Sodium alginate (1% by weight aqueous solution) 15.0
(6) Polyglyceryl monolaurate 1.0
(7) Alanine 0.01
(8) Proline 0.01
(9) Arginine 0.01
(10) Glutamic acid 0.01
(11) Glycine 0.01
(12) Macadamia nut oil fatty acid phytosteryl 3.0
(13) N-lauroyl-L-glutamic acid di (phytosteryl-2-octyldodecyl) 2.0
(14) Hardened palm oil 2.0
(15) Squalane (from olive) 1.0
(16) Behenyl alcohol 0.75
(17) Beeswax 1.0
(18) Jojoba oil 1.0
(19) 1,3-butylene glycol 10.0
(20) Extract 1 0.01
Production method: The aqueous phase components (1) to (11) are mixed and dissolved by heating at 75 ° C. On the other hand, the oil phase components (12) to (18) are mixed and dissolved by heating at 75 ° C. Next, the oil phase component is added to the aqueous phase component and preliminary emulsification is performed, followed by uniform emulsification with a homomixer. After cooling, add (19) at 50 ° C and (20) at 40 ° C and mix uniformly.

    The skin external preparations shown in Examples 4 to 7 were compositions having an anti-aging effect.

    Skin external preparations containing the five types of amino acids of the present invention (alanine, proline, arginine, glutamic acid, glycine, or salts thereof), and skin external preparations containing mokumaou extract are skins that exhibit a high anti-aging effect. It is useful as an external preparation.

Claims (3)

    A skin external preparation characterized by containing 5 types of amino acids.     The skin external preparation according to claim 1, wherein the five kinds of amino acids are alanine, proline, arginine, glutamic acid, glycine, or a salt thereof.     The skin external preparation according to claim 1, further comprising an extract of Casuarina Adans. In five kinds of amino acids.